UID:
almahu_9949251677102882
Umfang:
1 online resource (769 p.)
Ausgabe:
2nd ed. 2010.
ISBN:
9786612830549
,
9781282830547
,
1282830546
,
9783642011443
,
3642011446
Serie:
Springer Protocols Handbooks,
Inhalt:
Antibodies are indispensable tools for research, diagnosis, and therapy. Recombinant approaches allow the modification and improvement of nearly all antibody properties, such as affinity, valency, specificity, stability, serum half-life, effector functions, and immunogenicity. Antibody Engineering provides a comprehensive toolbox covering the well-established basics but also many exciting new techniques. The protocols reflect the latest "hands on" knowledge of key laboratories in this still fast-moving field. Newcomers will benefit from the proven step-by-step protocols, which include helpful practical advice; experienced antibody engineers will appreciate the new ideas and approaches. The book is an invaluable resource for all those engaged in antibody research and development.
Anmerkung:
Description based upon print version of record.
,
Cloning of the Antigen-binding Site from Hybridoma -- Cloning of Variable Domains from Mouse Hybridoma by PCR -- Coning Hybridoma cDNA by RACE -- Construction of scFv Fragments from Hybridoma or Spleen Cells by PCR Assembly -- Generation of Antibody Repertoires -- Mouse Immune Libraries for the Generation of ScFv Fragments Directed Against Human Cell Surface Antigens -- Human Antibody Gene Libraries -- Synthetic Antibody Libraries -- Immune Libraries from Nonhuman Primates (NHP) -- Generation of Rabbit Immune Libraries -- Selection of Antibody Fragments from Combinatorial Libraries -- Immunotube Selections -- Phage Display and Selection in Microtitre Plates -- Phage Display and Selections on Biotinylated Antigens -- Phage Display and Subtractive Selection on Cells -- Selection of Phage Antibody Libraries for Binding and Internalization into Mammalian Cells -- Improving Phage Display Throughput by Using Hyperphage, Miniaturized Titration and pVIII (g8p) ELISA -- Yeast Display and Selections -- The Generation of Transgenic Mice Expressing Human Antibody Repertoires -- Selection of Antibody Fragments by Means of the Filter-Sandwich Colony Screening Assay -- Semi-automated Magnetic Bead-Based Antibody Selection from Phage Display Libraries -- Engineering and Production of Immunoglobulins -- Aspects of Isotype Selection -- Generation of Heavy and Light Chains (Chimeric Antibodies) -- Humanising Antibodies by CDR Grafting -- Humanization by Resurfacing -- Human Antibodies by Guided Selection -- In Silico De-Immunization -- Affinity Maturation by Chain Shuffling and Site Directed Mutagenesis -- Affinity Maturation by Random Mutagenesis and Phage Display -- Engineering of the Fc Region for Improved PK (FcRn Interaction) -- Antibody-Dependent Enzyme Prodrug Therapy (ADEPT) -- Production of Recombinant Human IgG Antibodies in the Baculovirus Expression System -- Expression of IgA Molecules in Mammalian Cells -- Antibody Characterization -- Expression of Complete Antibodies in Transgenic Plants -- Expression ofFull Length Monoclonal Antibodies (mAb) in Algal Chloroplast -- Expression of IgG Antibodies in Mammalian Cells -- Protein A/G Chromatography -- Epitope Analysis Using Synthetic Peptide Repertoires Prepared by SPOT Synthesis Technology -- Epitope Mapping by Printed Peptide Libraries -- Antibody Epitope Mapping Using Yeast Display -- Size Exclusion Chromatography -- Structural Characterization of Antibodies by Mass Spectrometry -- Antibody Glycans Characterization -- Affinity Measurements by Competition ELISA -- Anti-Histidine Antibodies as Tools for Reversible Capturing of His-Tagged Fusion Proteins for Subsequent Binding Analysis -- Affinity Measurements Using Quartz Crystal Microbalance (QCM) -- Affinity Measurements with Radiolabeled Antibodies -- Neutralization Tests -- Functional Characterization of Antibodies Neutralizing Soluble Factors In Vitro and In Vivo -- Competitive ELISA -- Quantification of Human IgG and Related Fc Fusion Proteins by a Human IgG/Fc Capture ELISA -- Determination of Fc-Mediated Antibody-Effector Functions by Chromium Release Assay -- Binding Studies with Flow Cytometry.
,
English
Weitere Ausg.:
ISBN 9783642011436
Weitere Ausg.:
ISBN 3642011438
Sprache:
Englisch
DOI:
10.1007/978-3-642-01144-3
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